Hypusinated Eukaryotic Initiation Factor 5a Blockade In Cancer And Fibrosis: A Possible Novel Therapeutic Target For Leiomyosarcoma And Leiomyoma

 1,2 * Alessandro Zannotti

1 Department of Specialist and Odontostomatological Clinical Sciences,Ancona,Italy

2 Department of Experimental and Clinical Medicine, Università Politecnica delle Marche, Ancona,Italy

*Corresponding author: Alessandro Zannotti, Department of Specialist and Odontostomatological Clinical Sciences, UniversitàPolitecnica delle Marche, Ancona, Italy,Department of Experimental and Clinical Medicine, Ancona,Italy

Citation: Alessandro Zannotti(2022)Hypusinated Eukaryotic Initiation Factor 5a Blockade In Cancer And Fibrosis: A Possible Novel Therapeutic Target For Leiomyosarcoma And Leiomyoma.1(1)

Copyright: © 2022 Alessandro Zannotti,This is an open-access article distributed under the terms of the Creative Commons Attribution License, whichpermitsunrestricted use, distribution, andreproductioninanymedium, providedthe original author and source arecredited.

Received: 23 February 2022| Accepted: 20 March 2022|Published: 29 April 2022

Keywords: leiomyosarcoma; leiomyoma; ecm; fibronectin; eIF5A; hypusination; gc-7; cancer; fibrosis

Introduction

The myometrium is the intermedium layer of the uterus and it is localized between the perimetrium and the endometrium.The myometrium can transform itself into leiomyosarcoma, a very rare malign tumor that shows a high mortality [1]. Myometrium can transform itself also into leiomyoma, a benign pathology.

Leiomyoma, (uterine leiomyoma, fibroid or myoma) is the most common female pelvic benign tumor [2]. Even if it is not malignant, leiomyoma causes significant morbidity. Among the most important symptoms associated to leiomyoma, it is noteworthy to remember prolonged or heavy menstrual bleeding, pelvic pressure and also pelvic pain, last but not least reproductive dysfunction [3,4].

These two pathologies are very different, but they have a common origin. So, it is very important to find the differences as well as the common features they share. This could represent the starting point in order to find a possible novel therapeutic target for these two different and, at the same time similar, pathologies.

Anyway, the most important problem about the leiomyoma is the fact that its etiology remains still unknown [5]. It is considered a typical fibrotic disorder. In fact, the extracellular matrix (ECM) proteins—above all, collagen 1A1, versican and fibronectin—are upregulated in this benign pathology [6]. It was proposed that a dysregulated inflammation process could drive an excessive wound healing process that, in turn, may lead to the altered ECM [7,8] characterizing leiomyoma. Several cellular types act in the complex inflammatory response [9]. In particular, macrophages could play an important role in the dysregulated inflammation response whitin leiomyoma. In fact, something dysregulated in monocyte chemoattractant protein-1 (MCP-1) and granulocyte macrophage-colony-stimulating factor (GM-CSF) acting on macrophage proliferation, accumulation and infiltration could play a key role in the uncontrolled tissue repair and in the consequent pathological fibrosis that characterizes leiomyoma [10]. Furthermore, also something dysregulated in the molecules secreted by macrophages such as transforming growth factor-beta (TGF-β), activin A and tumor necrosis factor-alfa (TNF-α) could, in turn, contribute to the pathological fibrosis in leiomyoma [10]. In addition, the ECM seems to represent a reservoir for these kinds of molecules which are all molecules thought to be involved in the initiation and development of the leiomyoma [10].

In such a complicated pathology we have also to consider the existing interconnection between ECM, cellular membrane and inner cellular environment [11,14]. In fact, the cellular membrane is actively involved in processes such as nutrients and oxygen diffusion, the reception and the propagation of the signals for the cell adaptation to physiological and pathological environments [11,14]. About this question a clarifier example is represented by the results obtained in a recent study about the effects of Omega-3 fatty acids on the leiomyoma cells. It has been demonstrated that Omega-3 fatty acids can modulate the lipids profile [11], they can alter the cellular membrane architecture [11] and they can also downregulate the expression of genes involved in the mechanical signaling process [11] and in the lipids accumulation in the leiomyoma cells [11].

Since the interconnection between the ECM, the cellular membrane and the inner cellular environment has effects also on the gene expression, molecules such as translation factors could carry out an important role in the leiomyoma etiology.

Among the translation factors, in particular, eukaryotic initiation factor 5A (eIF5A) could play a key role in the establishment and the development of leiomyosarcoma and leiomyoma. In fact, it was demonstrated that this translation factor is involved in tumors, in cell proliferation [15, 16] and in the process of apoptosis [15]. In addition to this, it was observed that eIF5A modulates macrophage activation [17] and this may contribute to explain the eIF5A involvement in inflammation and fibrosis. In fact, it was reported that eIF5A could have an important role in the inflammation and in the fibrosis [18,19].

In addition to this, the translation factor eIF5A is the only protein containing hypusine residue in the human genome. The hypusine consists in a post-translational modification of the lysine residue to which a polyamine component from spermidine is conjugated [16]. This process is called hypusination [16] and it involves two different enzymes: the Deoxyhypusine synthase (DHPS) and the Deoxyhypusine hydroxylase (DOHH) [16].

Anyway, it was found that eIF5A can be synthetized in the hypusinated form and also in the not hypusinated one [16]. It was observed that the hypusinated eIF5A behaves like a translation initiating factor [16]. Furthermore, hypusinated eIF5A behaves also like translation elongation factor stimulating different types of mRNAs depending on the tissues [16]. It was demonstrated that the hypusinated eIF5A can also bind different mRNAs during the RNA export phase [16]. On the other hand, it was observed that the not hypusinated eIF5A does not develop these functions and it is not known if it can be involved in any other processes [16].

It was demonstrated that DOHH can be inhibited by ciclopirix and defiriprone [20] leading to the stop of the cell proliferation in the cervix cancer [20].

Besides this, it is known that the N1-guanyl-1,7-diamineheptane (GC-7) is the specific inhibitor of the DHPS [15]. The GC-7, as a spermidine analog, can bind the DHPS inhibiting the hypusination [15].

So, in this review it was explained the role of eIF5A translation factor and, within it, of its residue hypusine in both cancer and fibrosis. The aim of this review was to show how much this translation factor could represent a possible novel therapeutic target for leiomyosarcoma and leiomyoma acting directly on this translation factor or on the hypusination reaction.

The Role of Eif5a And The Effects Of Eif5a Blockade In Cancer And Fibrosis

It was demonstrated that eIF5A is one of the top four genes that are upregulated in the colon tumors [15].In addition to this, it was shown that eIF5A overexpression promotes epithelial-mesenchymal transition (EMT) and so, it leads to invasion and metastasis in hepatocellular carcinoma [21], bladder cancer [22], esophageal squamous cell carcinoma [23] and non-small cell lung cancer (NSCLC) [24].

These results were confirmed and enriched by those obtained by Meng et al. about human gastric cancer (GC). In fact, they demonstrated that elevated levels of eIF5A and its potential target metastasis-associated protein (MTA1) correlate with lymphovascular invasion and with more advanced stages of disease [25]. They also showed that there is a correlation between overexpression of eIF5A and its potential target MTA1 with disease progression [25]. These data agree with those that He et al.

Had previously obtained analyzing GC. In fact, they had shown that elevated levels of eIF5A correlate with poor prognosis in patients affected by GC [26]. In addition to this, He et al. had observed that there is an important correlation between elevated levels of eIF5A and advanced clinical stage also in NSCLC [26]. In addition, Yang et al. had found similar results in ovarian cancer [27].

On the other hand, acting directly on this translation factor or on the hypusination reaction, effects opposite to the ones just described were obtained. In fact, several studies agree with the fact that eIF5A knockdown impairs cell proliferation and also cell migration and invasion, while the eIF5A overexpression causes the opposite effects [28-30]. Moreover, Wei et al. showed that the downregulation of eIF5A impairs metastasis in xenograft studies [22].

In addition, it was demonstrated that tumorigenic potential in tumor cells and cancer cell growth are reduced after eIF5A knockdown, respectively in vivo and ex vivo [15]. In addition to this, it was reported in literature that not only eIF5A blocking performed by siRNA- or shRNA-directed eIF5A knockdown, but also performed by preventing the hypusination reaction through the GC-7 treatment causes the inhibition of tumor cell growth ex vivo and of tumorigenic potential in mouse xenograft models [15].

Furthermore, it was demonstrated that high levels of intracellular polyamines play a key role in the development of colorectal cancer (CRC) and that the ability of the difluoromethylornithine (DFMO) to inhibit tumor formation and progression by blocking the ornithine decarboxylase (ODC), that is in turn the first and rate limiting enzyme in the polyamine biosynthesis pathway, is only transient [31].

In this way, it is easy to understand that it is important to identify drugs that could stably interfere with these pathological processes.

Since it had been demostrated that eIF5A is overexpressed in CRC, that this feature is associated to poor prognosis [32] and that eIF5A is involved in the regulation of many polyamines pathways, such as cell proliferation, viability, migration, and autophagy [33-35], Coni et al. studied the effect of the GC-7 on hypusinated eIF5A in CRC. They showed that the eIF5A hypusination blocking by GC-7 leads to the inhibition of CRC cells growth in vitro and in vivo [36]. This finding was confirmed in mice, where GC-7 impairs the growth of CRC cells and polyps [36]. These results agree with those obtained by Fujimura et al. about KRAS-mutated pancreatic ductal adenocarcinoma cells [37]. Anyway, Coni provided a different mechanistic explanation for the ability of eIF5A in stimulating tumor progression. Coni’s results suggest that hypusinated eIF5A is not related to KRAS expression, instead, hypusinated eIF5A affects MYC protein expression. In particular, the inhibition of eIF5A hypusination impairs MYC protein levels and in this way, it reduces tumor growth [36]. Beyond the mechanism, the important thing is that both works show the importance of eIF5A hypusination in tumor progression. Moreover, the most remarkable aspect is that both works show that eIF5A hypusination blockage through the specific DHPS inhibitor GC-7 leads to the decrease of tumor progression.

In addition to this, it was reported that hypusinated eIF5A could play an important role in the inflammation and in the consequent fibrosis [18,19] that are two processes characterizing leiomyoma pathology [10]. In particular, it was demonstrated that polyamines and eIF5A hypusination modulate macrophage activation [17] that, in turn, was demonstrated to play a key role in the development of the fibrotic process in leiomyoma pathology [10]. In detail, Puleston et al. demonstrated, by inhibiting the polyamine-eIF5A-hypusine pathway, that hypusinated eIF5A is important for the oxidative phosphorylation (OXPHOS)-dependent macrophage alternative activation, since it was impaired after the blocking of the polyamine-eIF5A-hypusine pathway [17].

Maier et al. had already provided a proof that the inhibition of the hypusination reaction by GC-7 impaires the inflammation in β cells [38].

Moreover, it was reported that  inflammation and fibrosis, in leiomyoma pathology, lead to the upregulation of the ECM proteins and, among them, of the fibronectin [10]. In addition, this molecule was demonstrated to be involved also in invasion and metastasis development in cancer [39-47]. Thus, it is noteworthy to report the results obtained by Güth et al. about the hypusination of eIF5A and its involvement in the TGFβ/fibronectin-induced breast cancer metastasis. In fact, they demonstrated that the expression of the Pseudopodium-Enriched Atypical Kinase One (PEAK1) that is a cytoskeleton-associated kinase fundamental for non-canonical Transforming Growth Factor β (TGFβ) signaling and TGFβ/fibronectin-induced metastasis [48-50], is impaired by inhibition via GC-7 of DHPS-dependent eIF5A hypusination [51]. In addition to this, Güth et al. demonstrated that the inhibition of eIF5A hypusination by GC-7 impairs also the expression of E-Cadherin, breast cancer cell viability and, above all, TGFβ/fibronectin-induced PEAK1-dependent breast cancer metastasis [51]. On the other hand, they noticed that, in high-grade metastatic breast cancer cells, eIF5A hypusination is promoted by TGFβ stimulation [51].

These results highlight, one more time, the interconnection between eIF5A hypusination and metastasis and also their interconnection with TGFβ and fibronectin that in turn are molecules known to be connected with pathological fibrosis [10]. In addition to all this, it has already been mentioned the involvement of eIF5A also in inflammation and fibrosis.

Conclusion

In conclusion, evidences about the importance of fibronectin in fibrosis and metastasis development were reported.

In addition to this, evidences about the involvement of eIF5A in inflammation, in macrophage activation and in fibrosis that are all fundamental features for the initiation and development of benign leiomyoma were reported.

Furthermore, evidences about the involvement of eIF5A also in cancer and metastasis that is a fundamental feature of malign leiomyosarcoma were reported.

Moreover, this review related these findings to each other and to the fact that the eIF5A blockade, in particular, hindering eIF5A hypusination, leads to a decrease in tumor and fibrotic progression in several diseases.

All this may contribute to candidate eIF5A and, in particular, the inhibition of hypusination as therapeutic targets for both leiomyosarcoma and leiomyoma that are two different pathologies that, on the other hand, share some features.

Funding: This research received no external funding.

Conflicts of Interest: The author declares no conflict of interest.

References

1. Juhasz-Böss I, Gabriel L Bohle, RM Horn LC, Solomayer EF, Breitbach GP. (2018) Uterine Leiomyosarcoma. Oncol Res Treat 41(11): 680-686
2. Mathew RP Francis, S Jayaram V, Anvarsadath S. (2021) Uterine leiomyomas revisited with review of literature. Abdom Radiol (NY) 46(10): 4908-4926
3. Stewart EA. (2001) Uterine fibroids. Lancet 357(9252) 293-298.
4. Buttram VC, Jr Reiter RC. (1981) Uterine leiomyomata: etiology, symptomatology, and management. Fertil Steril 36(4) 433-445.
5. Islam MS, Protic O, Giannubilo SR, Toti P, Tranquilli AL et al. (2013)Uterine leiomyoma: available medical treatments and new possible therapeutic options. J Clin Endocrinol Metab 98(3) 921-934.
6. Gelse K, Pöschl E, Aigner T. (2003) Collagens--structure, function, and biosynthesis. Adv Drug Deliv Rev 55(12) 1531-1546.
7. Friedman AJ,Rein MS, Harrison-Atlas D,Garfield JM, Doubilet PM. (1990) A randomized, placebo-controlled, double-blind study evaluating leuprolide acetate depot treatment before myomectomy. Fertil Steril 52: (5)728-733.
8. Leppert PC, Catherino WH, Segars JH. (2006) A new hypothesis about the origin of uterine fibroids based on gene expression profiling with microarrays. Am J Obstet Gynecol 195 (2 ) 415-420.
9. Wynn TA. (2008) Cellular and molecular mechanisms of fibrosis. J Pathol 214(2) 199-210.
10. Zannotti A, Greco S, Pellegrino P, Giantomassi F, Delli Carpini G, et al. (2021) Macrophages and Immune Responses in Uterine Fibroids. Cells 10(5) 982.
11. Islam MS, Castellucci C, Fiorini R, Greco S, Gagliardi R et al. (2018) Omega-3 fatty acids modulate the lipid profile, membrane architecture, and gene expression of leiomyoma cells. J Cell Physiol 233(9) 7143-7156.
12. Protic O Islam, MS Greco S, Giannubilo SR, Lamanna P,Petraglia F et al.(2017) Activin A in Inflammation, Tissue Repair, and Fibrosis: Possible Role as Inflammatory and Fibrotic Mediator of Uterine Fibroid Development and Growth. Semin Reprod Med 35 (6) 499-509.
13. Ciarmela P, Islam MS, Reis FM,Gray PC, Bloise E, et al.(2011) Growth factors and myometrium: biological effects in uterine fibroid and possible clinical implications. Hum Reprod Update 17 (6) 772-790.
14. Islam MS, Ciavattini A, Petraglia F, Castellucci M, Ciarmela P. (2018) Extracellular matrix in uterine leiomyoma pathogenesis: a potential target for future therapeutics. Hum Reprod Update 24(1) 59-85.
15. Nakanishi S, Cleveland JL. (2016) Targeting the polyamine-hypusine circuit for the prevention and treatment of cancer. Amino Acids. 48(10) 2353-2362.
16. Caraglia M, Park MH ,Wolff EC, Marra M,Abbruzzese A. (2013) eIF5A isoforms and cancer: two brothers for two functions? Amino Acids 44(1) 103-109.
17. Puleston DJ, Buck MD,Klein Geltink RI, Kyle RL,Caputa G et al. (2019) Polyamines and eIF5A Hypusination Modulate Mitochondrial Respiration and Macrophage Activation. Cell Metab 30(2) 352-363.e8.
18. Kaiser A. (2012) Translational control of eIF5A in various diseases. Amino Acids 42: 679-684.
19. Pearce EJ, MacDonald AS. (2002) The immunobiology of schistosomiasis. Nat Rev Immunol 2(7) 499-511
20. Mémin E Hoque, M Jain MR, Heller DS, Li H, Cracchiolo B et al. (2014) Blocking eIF5A modification in cervical cancer cells alters the expression of cancer-related genes and suppresses cell proliferation. Cancer Res 74(2) 552-562.
21. Tang DJ, Dong SS, Ma NF, Xie D, Chen L. et al. (2010) Overexpression of eukaryotic initiation factor 5A2 enhances cell motility and promotes tumor metastasis in hepatocellular carcinoma. Hepatology 51(4) 1255-1263.
22. Wei JH,Cao JZ, Zhang D, Liao B, Zhong WM et al. (2014) EIF5A2 predicts outcome in localised invasive bladder cancer and promotes bladder cancer cell aggressiveness in vitro and in vivo. Br J Cancer 110(7) 1767-1777.
23. Li Y, Fu L,Li JB, Qin Y, Zeng TT et al. (2014) Increased expression of EIF5A2, via hypoxia or gene amplification, contributes to metastasis and angiogenesis of esophageal squamous cell carcinoma. Gastroenterology 146(7)1701-1713.
24. Xu G, Yu H, Shi X, Sun L, Zhou Q et al. (2014) Cisplatin sensitivity is enhanced in non-small cell lung cancer cells by regulating epithelial-mesenchymal transition through inhibition of eukaryotic translation initiation factor 5A2. BMC Pulm Med 14:174.
25. Meng QB, Kang WM, Yu JC, Liu YQ, Ma ZQ, et al. (2015) Overexpression of eukaryotic translation initiation factor 5A2 (EIF5A2) correlates with cell aggressiveness and poor survival in gastric cancer. PLoS One 10: 3 e0119229.
26. He LR ,Zhao HY, Li BK, Liu YH, Liu MZ et al. (2011) Overexpression of eIF5A-2 is an adverse prognostic marker of survival in stage I non-small cell lung cancer patients. Int J Cancer 129(1) 143-150.
27. Yang GF, Xie D, Liu JH, Luo JH, Li LJ et al. (2009) Expression and amplification of eIF-5A2 in human epithelial ovarian tumors and overexpression of EIF-5A2 is a new independent predictor of outcome in patients with ovarian carcinoma. Gynecol Oncol 112 (2) 314-318.
28. Zhu W, Cai MY, Tong ZT, Dong SS, Mai SJ et al. (2012) Overexpression of EIF5A2 promotes colorectal carcinoma cell aggressiveness by upregulating MTA1 through C-myc to induce epithelial-mesenchymaltransition. Gut 61(4) 562–575.
29. Khosravi S, Wong RP, Ardekani G, Zhang G Martinka M et al. (2014) Role of EIF5A2, a downstream target of Akt, in promoting melanoma cell invasion. Br J Cancer 110(2) 399-408.
30. Zender L,Xue W ,Zuber J,Semighini CP,Krasnitz A et al. (2008) An oncogenomics-based in vivo RNAi screen identifies tumor suppressors in liver cancer. Cell 135(5) 852-864.
31. LoGiudice N, Le L, Abuan I,Leizorek Y, Roberts SC. (2018) Alpha-Difluoromethylornithine, an Irreversible Inhibitor of Polyamine Biosynthesis, as a Therapeutic Strategy against Hyperproliferative and Infectious Diseases. Med Sci 6: 1- 12.
32. Tunca B, Tezcan G, Cecener G, Egeli U, Zorluoglu A et al. (2013) Overexpression of CK20, MAP3K8 and EIF5A correlates with poor prognosis in early-onset colorectal cancer patients. J Cancer Res Clin Oncol 139 (4) 691-702.
33. Turpaev KT. (2018) Translation Factor eIF5A, Modification with Hypusine and Role in Regulation of Gene Expression. eIF5A as a Target for Pharmacological Interventions. Biochemistry 83(8) 863-873.
34. Lubas M, Harder LM, Kumsta C, Tiessen I,Hansen M et al. (2018) eIF5A is required for autophagy by mediating ATG3 translation. EMBO Rep 19: 6 e46072.
35. Zhang H, Alsaleh G, Feltham J, Sun Y, Napolitano G et al. (2019) Polyamines Control eIF5A Hypusination, TFEB Translation, and Autophagy to Reverse B Cell Senescence. Mol Cell 76: 1 110-125.e9.
36. Coni S, Serrao SM, Yurtsever ZN, Di Magno L, Bordone R et al. (2020) Blockade of EIF5A hypusination limits colorectal cancer growth by inhibiting MYC elongation. Cell Death Dis 11: (12) 1045.
37. Fujimura K, Wang H, Watson F, Klemke RL. (2018) KRAS Oncoprotein Expression Is Regulated by a Self-Governing eIF5A-PEAK1 Feed-Forward Regulatory Loop. Cancer Res 78: (6) 1444-1456.
38. Maier B, Ogihara T, Trace AP, Tersey SA, Robbins RD et al. (2010) The unique hypusine modification of eIF5A promotes islet beta cell inflammation and dysfunction in mice. J Clin Invest 120 (6) :2156-2170.
39. Indra I, Beningo KA. (2011) An in vitro correlation of metastatic capacity, substrate rigidity, and ECM composition. J Cell Biochem 112(11) :3151-3158.
40. Lin TC, Yang CH, Cheng LH, Chang WT, Lin YR. (2019) Fibronectin in Cancer: Friend or Foe. Cells 9: 1 27.
41. D'Ardenne AJ, Kirkpatrick P, Sykes BC. (1984) Distribution of laminin, fibronectin, and interstitial collagen type III in soft tissue tumours. J Clin Pathol 37(8) 895-904.
42. Itagaki K, Sasada M, Miyazaki S, Iyoda T, Imaizumi T, et al. (2020) Exposure of the cryptic de-adhesive site FNIII14 in fibronectin molecule and its binding to membrane-type eEF1A induce migration and invasion of cancer cells via β1-integrin inactivation. Am J Cancer Res 10: (11): 3990-4004.
43. Wu S Zheng, Xing Q, Dong X, Wang Y, You Y et al. (2018) Matrix stiffness-upregulated LOXL2 promotes fibronectin production, MMP9 and CXCL12 expression and BMDCs recruitment to assist pre-metastatic niche formation. J Exp Clin Cancer Res 37(1) 99.
44. Lv ZD, Na D, Liu FN, Du ZM, Sun Z et al. (2010) Induction of gastric cancer cell adhesion through transforming growth factor-beta1-mediated peritoneal fibrosis. J Exp Clin Cancer Res 29(1) 139.
45. Ahmed N Riley, Rice C, Quinn GM. (2005) Role of integrin receptors for fibronectin, collagen and laminin in the regulation of ovarian carcinoma functions in response to a matrix microenvironment. Clin Exp Metastasis 22(5) 391-402.
46. Barbazán J ,Matic Vignjevic D. (2019) Cancer associated fibroblasts: is the force the path to the dark side? Curr Opin Cell Biol 56: 71-79.
47. Theocharis AD, Manou D, Karamanos NK. (2019) The extracellular matrix as a multitasking player in disease. FEBS J 286: 15 2830-2869.
48. Agajanian M, Campeau A, Hoover M, Hou A, Brambilla D et al. 2015 PEAK1 Acts as a Molecular Switch to Regulate Context-Dependent TGFβ Responses in Breast Cancer. PLoS One 10: 8 e0135748.
49. Agajanian M, Runa F Kelber JA. (2015) Identification of a PEAK1/ZEB1 signaling axis during TGFβ/fibronectin-induced EMT in breast cancer. Biochem Biophys Res Commun 465(3) 606-612.
50. Yeh HW, Lee SS, Chang CY, Lang YD, Jou YS. (2019) A New Switch for TGFβ in Cancer. Cancer Res 79(15) 3797-3805.
51. Güth R, Adamian Y, Geller C, Molnar J,Maddela J et al. (2019) DHPS-dependent hypusination of eIF5A1/2 is necessary for TGFβ/fibronectin-induced breast cancer metastasis and associates with prognostically unfavorable genomic alterations in TP53. Biochem Biophys Res Commun 519 (4) 838-845.